Method of applying adenovirus vector mediation ribose nucleic acid (RNA) jamming technology to silence sterol regulatory element binding protein 1

Abstract

The invention discloses a method of applying an adenovirus vector mediation ribose nucleic acid (RNA) jamming technology to silence a sterol regulatory element binding protein 1. The method of applying the adenovirus vector mediation RNA jamming technology to silence the sterol regulatory element binding protein 1 comprises the following steps: A1, cloning a Qinchuan cattle sterol regulatory element binding protein 1 (SREBP1); A2, constructing a psiCHECK-II-SREBP1 expression vector; A3, designing and synthesizing a target SREBP1 gene sh RNA; A4, constructing a pENTR / CMV-GFP / U6-sh RNA expression vector; A5, determining and screening interference efficiency of sh RNA; A6, constructing a recombinant adenovirous vector; and A7, carrying out adenovirus packaging, amplification and titre determination. The method of applying the adenovirus vector mediation RNA jamming technology to silence the sterol regulatory element binding protein 1 is an effective method of applying the RNA jamming technology to research functions of the Qinchuan cattle SREBP1 gene.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for silencing sterol regulatory element binding protein gene 1 (SREBP1) using adenovirus vector-mediated RNA interference technology Background technique [0002] Sterol regulatory element binding proteins (Sterol regμlatory element binding proteins: SREBPs) are membrane-bound proteins that belong to the family of basic helix-loop-helix leucine zipper transcription factors, which mainly promote glycolysis and lipogenesis in It plays an important role in energy balance (Eberle et al., 2004; Shimomura et al., 1997; Wang et al., 1994). Studies have shown that the SREBPs family is formed by binding to the SRE (5' -TCACNCCAC-3') sequence to regulate gene transcription (Shimano, 2001). SREBP-1c can promote the expression of PPARγ and the production of endogenous PPARγ, and cooperate with PPARγ and CCAAT enhancer binding protein (CCAAT enhancer binding protein, C / EBPs)...

AI Technical Summary

Problems solved by technology

In most cases, shRNA designed for a specific gene cannot achieve the effect of silencing the gene (Elbashir et al., 2002a; Holen et al., 2002; KapadiaBrideau-Andersen and Chisari, 2003; Lee et al., 2002; YuDeRuiter and Turner, 2002), generally only 1 / 5 can effectively silence the target gene (KapadiaBrideau-Andersen and Chisari, 2003; McManus et al., 2002)

Images